Quantum dot loaded immunomicelles for tumor imaging

<p>Abstract</p> <p>Background</p> <p>Optical imaging is a promising method for the detection of tumors in animals, with speed and minimal invasiveness. We have previously developed a lipid coated quantum dot system that doubles the fluorescence of PEG-grafted quantum dots at half the dose. Here, we describe a tumor-targeted near infrared imaging agent composed of cancer-specific monoclonal anti-nucleosome antibody 2C5, coupled to quantum dot (QD)-containing polymeric micelles, prepared from a polyethylene glycol/phosphatidylethanolamine (PEG-PE) conjugate. Its production is simple and involves no special equipment. Its imaging potential is great since the fluorescence intensity in the tumor is twofold that of non-targeted QD-loaded PEG-PE micelles at one hour after injection.</p> <p>Methods</p> <p>Para-nitrophenol-containing (5%) PEG-PE quantum dot micelles were produced by the thin layer method. Following hydration, 2C5 antibody was attached to the PEG-PE micelles and the QD-micelles were purified using dialysis. 4T1 breast tumors were inoculated subcutaneously in the flank of the animals. A lung pseudometastatic B16F10 melanoma model was developed using tail vein injection. The contrast agents were injected via the tail vein and mice were depilated, anesthetized and imaged on a Kodak Image Station. Images were taken at one, two, and four hours and analyzed using a methodology that produces normalized signal-to-noise data. This allowed for the comparison between different subjects and time points. For the pseudometastatic model, lungs were removed and imaged <it>ex vivo </it>at one and twenty four hours.</p> <p>Results</p> <p>The contrast agent signal intensity at the tumor was double that of the passively targeted QD-micelles with equally fast and sharply contrasted images. With the side views of the animals only tumor is visible, while in the dorsal view internal organs including liver and kidney are visible. <it>Ex vivo </it>results demonstrated that the agent detects melanoma nodes in a lung pseudometastatic model after a 24 hours wash-out period, while at one hour, only a uniform signal is detected.</p> <p>Conclusions</p> <p>The targeted agent produces ultrabright tumor images and double the fluorescence intensity, as rapidly and at the same low dose as the passively targeted agents. It represents a development that may potentially serve to enhance early detection for metastases.</p

Istraživanje mehanizma toksičnosti anilina u eritrocitima

Strategies for the use of bio-indicators in the prediction of environmental damage should include mechanistic research. This study involves the relationship between the chemical structure and hemotoxic markers of aniline and its halogenated analogs. Aniline-induced methemoglobinemia, loss of circulating blood cells, blood stability, glutathione depletion and membrane cytoskeletal changes were assessed following exposure to phenylhydroxylamine (PHA), para-fluoro-, para-bromo-, and para-iodo in male Sprague-Dawley rats. Methemoglobin was determined spectrophotometrically at 635 nm. Erythrocyte depletion was investigated by loss of radioactivity in chromium-labeled red blood cells in vivo. Membrane proteins were analyzed by SDS-PAGE using red blood ghost cells treated with various aniline analogs. Results showed dose- and time-dependent changes in the induction of methemoglobin of up to 78 % with para-bromo PHA and 75 % with para-iodo PHA compared to 3 % to 5 % in control. Treated animals lost up to three times more blood from circulation compared to control within 14 days after treatment. Erythrocytes were more stable in buffer solution than in para-iodo-treated cells. Depletion of reduced glutathione in PHA and para-iodo-PHA treated red cells was also observed. Analysis of red cell skeletal membrane treated with para-iodo-PHA showed that protein band 2.1 became broader and band 2.2 diminished completely in some treatments. Dose- and time-dependent changes suggested the use of hemotoxic endpoints as potential biomarkers for assessing chemical and drug safetyStrategije primjene biopokazatelja za predviđanje štete u okolišu trebaju u obzir uzeti istraživanja mehanizama djelovanja. Ovo istraživanje propituje odnos između kemijske strukture i hemotoksičnih pokazatelja djelovanja anilina i njegovh halogeniranih analoga. Nakon izlaganja mužjaka štakora soja Sprague-Dawley para-fluoro-, para-bromo- i para-jodofenilhidroksilaminu, utvrđena je methemoglobinemija uzrokovana anilinom te pad broja krvnih stanica u krvotoku i stabilnosti krvi, gubitak glutationa i promjene na membrani stanice. Methemoglobin je određivan spektrofotometrijski na 635 nm. Pad broja eritrocita mjeren je in vivo s pomoću eritrocita obilježenih radioaktivnim kromom. Membranske su bjelančevine analizirane s pomoću SDS-PAGE, rabeći eritrocite bez hemoglobina (engl. ghost cells) kojima su dodani različiti analozi anilina. Nalazi upućuju na promjene indukcije methemoglobina ovisno o dozi i vremenu djelovanja do 78 % s para-bromo-fenilhidroksilaminom te do 75 % s para-jodofenilhidroksilaminom u usporedbi s 3 % do 5 % u kontrolnih uzoraka. U razdoblju od 14 dana nakon tretiranja izložene životinje izgubile su tri puta više krvi iz krvotoka od kontrolnih. Eritrociti su bili stabilniji u puferskoj otopini negoli u stanicama kojima je dodan para-jodofenilhidroksilamin. Zamijećen je i pad glutationa u eritrocitima kojima je dodan fenilhidroksilamin odnosno para-jodofenilhidroksilamin. Analizom membrane eritrocita kojima je dodan para-jodofenilhidroksilamin zamijećeno je da se u pojedinih obrada raširila proteinska vrpca 2.1, a potpuno smanjila proteinska vrpca 2.2. Zamijećene promjene uvjetovane dozom i vremenom upućuju na primjenu hemotoksičnih parametara kao mogućih biopokazatelja u procjeni sigurnosti lijeka odnosno kemikalije

Correlation between the out-of-Plane Components of Magnetizability and Central Magnetic Shielding in Unsaturated Cyclic Molecules

A simple classical model of magnetic-field induced pi-electron flow is discussed, showing that the contribution to the sigma(parallel to) out-of-plane component of the virtual magnetic shielding provided by pi-ring currents, at points P along the C axis of cyclic planar unsaturated hydrocarbons C H with D-nh symmetry, in the presence of a magnetic field B-ext at right angles to the a plane, is, with good approximation, connected with the pi-electron contribution to the out-of-plane component of the magnetizability, xi(parallel to). The relationship is sigma(parallel to) (h) = (mu(0)/2 pi) (s(2) + h(2))(-3/2) xi(parallel to), where s is the distance of a C nucleus from the center of the carbon ring, and h is the distance of P from a. The ring current susceptibility, that is, the strength of the pi currents, expressed in nA/T (nano ampere per tesla) within the SI system of units, is given by partial derivative I/partial derivative B-ext = xi(parallel to)(pi s(2)), which can be used as a reliable virtual measure of magnetotropicity and relative pi-electron mobility in isoelectronic systems. Criteria for the practicality of the proposed ring current model are discussed

Structural and mechanistic insights into 5-lipoxygenase inhibition by natural products

© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc. Leukotrienes (LT) are lipid mediators of the inflammatory response that are linked to asthma and atherosclerosis. LT biosynthesis is initiated by 5-lipoxygenase (5-LOX) with the assistance of the substrate-binding 5-LOX-activating protein at the nuclear membrane. Here, we contrast the structural and functional consequences of the binding of two natural product inhibitors of 5-LOX. The redox-type inhibitor nordihydroguaiaretic acid (NDGA) is lodged in the 5-LOX active site, now fully exposed by disordering of the helix that caps it in the apo-enzyme. In contrast, the allosteric inhibitor 3-acetyl-11-keto-beta-boswellic acid (AKBA) from frankincense wedges between the membrane-binding and catalytic domains of 5-LOX, some 30 Å from the catalytic iron. While enzyme inhibition by NDGA is robust, AKBA promotes a shift in the regiospecificity, evident in human embryonic kidney 293 cells and in primary immune cells expressing 5-LOX. Our results suggest a new approach to isoform-specific 5-LOX inhibitor development through exploitation of an allosteric site in 5-LOX. [Figure not available: see fulltext.]